EP0357611A1 - Verfahren zum nachweis des karzinogenen menschlichen papillomavirus. - Google Patents

Verfahren zum nachweis des karzinogenen menschlichen papillomavirus.

Info

Publication number
EP0357611A1
EP0357611A1 EP88902077A EP88902077A EP0357611A1 EP 0357611 A1 EP0357611 A1 EP 0357611A1 EP 88902077 A EP88902077 A EP 88902077A EP 88902077 A EP88902077 A EP 88902077A EP 0357611 A1 EP0357611 A1 EP 0357611A1
Authority
EP
European Patent Office
Prior art keywords
dna
hpv
hpv16
lane
polymerase
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
EP88902077A
Other languages
English (en)
French (fr)
Other versions
EP0357611A4 (en
EP0357611B1 (de
Inventor
Brian James Morris
Brian Nightingale
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Sydney
Original Assignee
University of Sydney
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=3772035&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=EP0357611(A1) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by University of Sydney filed Critical University of Sydney
Publication of EP0357611A1 publication Critical patent/EP0357611A1/de
Publication of EP0357611A4 publication Critical patent/EP0357611A4/en
Application granted granted Critical
Publication of EP0357611B1 publication Critical patent/EP0357611B1/de
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
    • C12Q1/701—Specific hybridization probes
    • C12Q1/708—Specific hybridization probes for papilloma

Definitions

  • This invention relates to a method for the specific detection of the DNA of papillomaviruses in clinical samples.
  • the test aims to differentiate, in the shortest possible time, whether cells from the anogenital region contain types of papillomavirus that are associated with cancer or whether they contain types of papillomavirus that are generally associated with benign lesions.
  • Cancer of the cervix is the most common cancer in women ( ⁇ 25% of all female cancer). Moreover, the incidence is increasing in younger women. Indeed, approximately 2% of routine cervical smears show abnormal cytology, indicating an epidemic. Such an epidemic is current in many western and developing countries. Sexual activity appears to be an important predisposing factor in the epidemiology of carcinogenesis and precancerous lesions. An early age of sexual intercourse and multiplicity of sexual partners are associated statistically with an increased risk of malignancy [Harris et al, Br. J. Cancer 42: 359-63, 1980].
  • the new techniques in molecular biology can be utilised to bypass such problems and provide more objective information.
  • nucleic acid hybridization techniques the viral DNA can be identified directly and at an earlier stage of infection. Indeed, using these approaches, HPV types have been found in both benign and premalignant lesions.
  • HPV6 de Villiers etal., J. Virol. 40: 932-5, 1981
  • HPV11 Gassman etal, Proc. Natl. Acad. Sci.
  • High-risk HPV16, HPV18, HPV31, HPV33, HPV35, HPV39, HPV43 and HPV44.
  • the frequency of occurrence of the higher risk types is in decreasing order.
  • HPV16 is most common (45-60%)
  • HPV18 is next most common (20-30%)
  • the others are rarer, the last 4 being discovered only recently and reported in 1986 (total frequency for all of these rarer types is, collectively, ⁇ 15%).
  • Other rarer types are likely to be discovered in due course.
  • the integration pattern usually interupts or deletes specific regions of the HPV16 or 18 DNA, but consistently leaves intact the E6 and E7 openreading frames (ORFs) ([Pater and Pater, Virology 145: 313-8, 1985], which continue to express, at least in cell lines derived from cervical carcinomas [Smotkin and Wettstein, Proc. Natl Acad. Sci. USA 83: 4680-4, 1986; Androphy et al, EMBO J. 6: 989-92, 1987; Baker et al, J. Virol 61: 962-71, 1987; Takebeet al, Biochem. Biophys. Res. Commun. 143: 837-44, 1987].
  • ORFs E6 and E7 openreading frames
  • HPV6b The sequence of HPV6b is given in Schwartz et al, EMBO J. 2: 2361-8, 1983.
  • the sequence of HPV11 is given in Dartmann et al, Virology 151: 124-30, 1986.
  • the sequence of HPV16 is given in Seedorf et al, Virology 145: 181-5, 1985.
  • the sequence of HPV18 is given in Matlashewski et al, J. Gen. Virol 67: 1909-16, 1986.
  • the sequence of HPV33 is given in Cole and Streeck, J. Virol 58: 991-5, 1986.
  • DNA is double-stranded. Each strand of DNA is a complementary 'mirror image' of the other. The DNA strands are held together by hydrogen bonding.
  • Our techniques for detecting viral DNAs are based on the ability of the unique sequence of nucleotides in a DNA strand to bond with ('hybridize') to a sequence complementary to it.
  • a DNA sequence for all or a unique part of a papillomavirus type it is possible to use this as a 'homing probe' in order to detect the virus in a sample of cervical cells from a patient.
  • the DNA for use as probe is labelled either with a radioactive isotope or nonradioactive label so that it can be detected later.
  • To increase the sensitivity of the test we have utilized a method for amplification of the HPV DNA sequences in the sample.
  • PCR PCR
  • a typical amplification factor is ⁇ 250,000 copies starting from one copy of viral DNA.
  • Such an approach not only increases sensitivity, but fulfills requirements of specificity, speed, simplicity, and amenabihty to nonradioactive detection methods expected of a more versatile testing procedure. It is also amenable to assembly as a kit and to automation, both of which we have accomplished.
  • the PCR technique is described in papers that deal with prenatal diagnostic testing for specific genetic abnormalities [Sakii et al, Science 230: 1350-4,
  • Patent Application AU-A-55323/86 "Amplification and Detection of Target Nucleic Acid by
  • a small, unique portion of the HPV DNA sequence is amplified by the PCR procedure.
  • a region in the E6 region has been chosen.
  • any other region in the viral DNA may also be chosen.
  • the PCR step requires two ⁇ 20mer oligonucleotide primers that flank the region to be amplified.
  • One primer is complementary to the (+)-strand of a region of the DNA and the other is complementary to the (-)-strand.
  • the annealing of primer to the ( +)-strand of denatured sample viral DNA, followed by extension with, e.g., the Klenow fragment o ⁇ Escherichia coli DNA polymerase, or other enzymes that carry out a similar reaction, and deoxynucleotide triphosphates results in the synthesis of a (-)-strand fragment containing a 'target' sequence residing between the hybridization sites of the primer.
  • a similar reaction occurs with the other primer, creating a new (+)-strand.
  • the principle of the method is shown in the following diagram.
  • DNA strands are themselves templates for the PCR primers, repeated cycles of denaturation, primer annealing, and extension result in the exponential accumulation of the ⁇ 100-200 bp region defined by the primers.
  • the specific DNA is detected. Various means are possible for doing this. The amount of DNA produced may be sufficient for direct visualization after electrophoresis on a gel and staining.
  • the present invention provides a method of detection of carcinogenic human papillomavirus HPV16 and/or HPV18 which comprises
  • step (ii) adding oligonucleotide primers defining each end of said characteristic DNA portion, (iii) cooling to aUow the primers to anneal to the dissociated DNA strands, (iv) adding a DNA polymerase, (v) at the cooled temperature allowing formation of DNA complementary to each strand of said characteristic DNA portion, (vi) heating to dissociation temperature, and repeating steps (iii) to (vi), optionally omitting step (iv) where a heat stable DNA polymerase is used.
  • the invention also extends to the specific oligonucleotide primers used, and to specific labelled oligonucleotide probes, as described hereinafter.
  • Viral DNA is amplified by a polymerase chain reaction.
  • Samples are applied to a charged nylon membrane using a dot manifold, along with appropriate standards and controls.
  • Filter is hybridized with labelled mixed viral DNA probes, one mixture containing probes for the 'high risk' (potentially carcinogenic) and the other probes for the 'low risk' HPVs.
  • oligonucleotides For each HPV type or category two synthetic oligonucleotides are synthesized for use as primers in the extension and amplification step of PCR. These oligonucleotides correspond to
  • DNA flanking a specific "target" sequence of interest this being a sequence of DNA in the viral genome that is unique for HPV and differs from one HPV type or category ('high risk' vs low risk') to the next.
  • target sequence of interest a sequence of DNA in the viral genome that is unique for HPV and differs from one HPV type or category ('high risk' vs low risk') to the next.
  • oligonucleotides described below to be suitable; however, other appropriate sequences can also be chosen for the specific viral types indicated or for other types of papillomavirus.
  • Suitable target sequence has been chosen within the E6 open reading frame of the viral genome.
  • Suitable DNA sequences flanking this target sequence for use as primers are as follows (where position number refers to the nucleotide sequence of the viral genome) and are the same for each of these HPVs:
  • HPV6/11 primer 1 5' ATGCCTCCACGTCTGCAAC 3'
  • primers have complementary sequences on HPV6 and HPV11. They differ from sequences in the genomes of HPV16, HPV18 and HPV33, where separate primers flanking a chosen DNA target sequence are synthesized for these, together with oligonucleotides suitable for use as probes, using the same principal as described above and below, respectively, for HPV6 and HPV11.
  • High risk HPVs For HPV16 and HPV 33 suitable sequences flanking a chosen target sequence are as follows:
  • HPV16/33 primer 1 5' TGAGGTATATGACTTTGCTTTT 3'
  • HPV16/33 primer 2 3 AATTAATCCACATAAT 5'
  • HPV18 primer 1 5 TGTCATAACCrrGAATGTCT 3' POSITION: 428 437
  • HPV18 primer 2 3' AAATGTATAGATTTTATTC 5'
  • the sample may then be divided and probed for each category of HPV types (high risk vs low risk).
  • Synthesize oligonucleotide (target) sequence for use as probe This corresponds to a region between the two primers, and is identical for each of the viral types HPV6 and HPV11, but differs, of course, from any other viral or other DNA sequence. These sequences are as follows:
  • HPV6/11 target oligonucleotide 5' GCAAGACGTTTAATCT 3'
  • oligonucleotide (target) sequence is synthesized for use as a probe:
  • HPV16/33 target oligonucleotide 5' GTGAGTATAGACATTAT 3'
  • target sequence is synthesized for use as a probe:
  • HPV18 target oligonucleotide 5' GATTTATTTGTGGTGTATAGA 3,
  • [ ⁇ - 32 P]dATP (sp. act 3000 Ci/mmol) 100 pmoles (i.e., 30 ⁇ l of a 10 mCi/ml solution)
  • nonradioactive labelling of oligonucleotides for use as hybridization probes are more appropriate for use in a HPV detection kit as they generally have acceptably long half-lives and avoid dangers from and the need for a licence for use of radioactivity.
  • Alkaline phosphatase labelled oligonucleotides of any sequence specified are available made-to-order from BRESA.
  • Lysis buffer 50 mM NaCl, 50 mM Tris, pH 7.5, 0.5% SDS, 10 mM EDTA.
  • Deproteinization reagent 1 phenol/chloroform/isoamyl alcohol (25:24:1, v/v)
  • Deproteinization reagent 2 chloroform/isoamyl alcohol (24:1, v/v)
  • DNA polymerase solution 1 U/ ⁇ l of DNA polymerase (Klenow fragment)
  • Prehybridization solution 6 x SSC, 25 x Denhardt's solution, 0.5% sodium dodecyl sulphate.
  • Hybridization solution 6 x SSC, 1 x Denhardt's solution and 0.5% SDS, with 100 ng/ml alkaline phosphatase linked oligonucleotide probe.
  • Wash solution 3 1 M NaCl, 0.1 M Tris.HCl, pH 9.5, 5 mM MgCl 2
  • Colour reagent solution 0.33 mg/ml nitroblue tetrazollium (NBT), 0.17 mg/ml 5-bromo-4-chloro-3-indoyl phosphate (BCIP), 0.33% v/v dimethylformamide in 0.1 M Tris.HCl, pH 9.5, 0.1M NaCl, 5mM Mg Cl 2
  • TE 10 mM Tris.HCl, pH 8.0, 1 mM EDTA
  • Denhardt's solution 5 g Ficoll, 5 g polyvinylpyrrolidone, 5 g bovine serum albumin, made up to
  • the scrape tissue may be kept at 4°C for 2-3 days or frozen at -20°C for longer term storage.
  • a DNA polymerase that is resistant to 95°C is used instead. Use of this enzyme facilitates the test to some extent and, most importantly, reduces costs to -20%.
  • Example(s) positive for HPV16 or HPV33 show as a discrete band of approximately 200 bp.
  • Samples positive for HPV6 or HPV11 show as a discrete band of approximately 120 bp.
  • Samples positive for HPV6/11 and HPV16/33 show as discrete 200 bp and 120 bp bands.
  • Probing membrane with alkaline phosphatase-linked oligonucleotide probe i) Prehybridization - Place membrane in plastic bag provided and add 10 ml of prehybridization solution. Seal bag and incubate at 30°C for 40 min with agitation. ii) Hybridization - Empty contents of bag and replace with 10 ml of hybridization solution. Reseal plastic bag and incubate at 30°C for at 40 mi ⁇ with agitation. iii) Washing - Remove membrane from bag and place in a dish containing 500 ml of wash solution 1 preheated to 37°C. Agitate for 10 minutes at this temperature.
  • the membrane is prehybridized for at least 1 h at 30°C. Several approaches can be used for this.
  • carrier DNA e.g., from herring sperm
  • Hybridization Pour off prehybridization mixture and replace with hybridization mixture, which is the same except for the addition of radioactively labelled viral DNA (1 ng/ml hybridization mixture).
  • the membrane is blotted with filter paper, but not allowed to dry, and while still moist, is taped to a sheet of 3MM Whatman paper, then covered with cling wrap. 2. This is applied to a suitable x-ray film (e.g., Kodak XR-5) in a dark-room and sealed in an autoradiography cassette with two intensifying screens (DuPont).
  • a suitable x-ray film e.g., Kodak XR-5
  • DuPont two intensifying screens
  • lysis buffer by adding 1 ml distilled water to lysis buffer powder and then mixing it with the enzyme powder in vial.
  • PCR polymerase chain reaction
  • Results have indicated the importance of direct HPV detection in patient evaluation. For example, many patients that had normal cytology of cervical cells were found to be infected with the carcinogenic varieties of HPV. This may reflect the fact that our test can detect infection at an earlier stage, perhaps even before the virus has had a chance to cause a morphological change in the cell. Moreover, the lesions associated with the potentially carcinogenic varieties of HPV are often flat, rather than the large obvious condylamas caused by the more benign varieties, and are therefore not as easily seen in the clinic.
  • HPV16/33 primers 4 ⁇ l of each dNTP (50 mM stock). 23 rounds of: 98°C for 2 min, then 37°C for 2 min, add Klenow, then 37°C for 2 min. Electrophoresis on 2% agarose gel. Ethidium bromide stained gel is shown in Figure 4. Lanes 1 and 3 are DNA size markers, viz. pBR322 cut with Hpa ⁇ l and SPP-1, respectively. A single DNA band of the expected size (-200 base pairs) can be seen in lane 2.
  • Lane 2 blank Lane 3 - 100 ng HPV33 cut with BglI/PstI
  • Hybridization in lane 1 shows that specimen Gap289 contained HPV 16.
  • Figure 9 depicts the stained gel result.
  • Figure 10 shows the result of radioactive target oligo probing.
  • Figure 1 1 shows the result of alkaline phosphatase target oligo probing.
  • a dark band of hybridization can be seen at the correct position in lane 1 for the patient specimen 's PCR products and bands in lanes 2 and 3 corresponding to the position of the whole virus. This confirms the presence of HPV 16 in the cervical scrape.
  • Lane 3 - -500 ng Hs biopsy Lane 4 - -500 ng Ls biopsy
  • HPV6/1 1 band can be seen in lane 3.
  • Figure 18 a single band of hybridization to the -200 bp HPV16/33 amplification products can be seen in each of lanes 6-3.
  • Figure 19 shows the result of a 1.5 h exposure to the x-ray film: hybridization can be seen to the ⁇ 200 bp HPV 16/33 amplification products.
  • Figure 20 is a dot blot probed with radioactive target oligo probe (Right: diagram of position of samples on dot blot; Left: result of hybridization).
  • HP V 16/33 amplification products can be seen for the biopsy (lanes 2-4) and the anal scrape (lane 5).
  • the lanes are as follows: Lane 1 - pBR322 cut with Hpall Lane 2 - Gap289 (1:10 3 ) PCR (HPV16/33 primers only) Lane 3 - Gap289 (1:10 6 ) PCR ( HPV16/33 primers only) Lane 4 - Gap289 (1:10 9 ) PCR ( HPV16/33 primers only) Lane 5 - Anal scrape Gap402 PCR (HPV6/11 and HP V16/33 primers) Lane 6 - Bs wart PCR (HPV6/11 and HPV16/33 primers) Lane 7 - Cervical scrape 11912 ( HPV 16/33 primers only) Lane 8 - Vaginal scrape 11912 ( HPV16/33 primers only)
  • Lane 1 Bacteriophage ⁇ cut with HindIII/EcoRI Lane 2 - 5 ⁇ g Bs wart DNA, BglII/BamHI Lane 3 - 5 ⁇ g Gap289, BglII/BamHI Lane 4 - HPV16/pBR322, BamHI Lane 5 - HPV33/plink, Bglll Lane 6 - SPP-1
  • Lane 1 - HPV 16 insert PCR products HPV 16/33 primers.
  • HPV 18 PCR products of -100 bp can be seen in lane 2.
  • lane 1 are the HPV 16 PCR products of ⁇ 200 bp.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Virology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
EP88902077A 1987-02-26 1988-02-24 Verfahren zum nachweis des karzinogenen menschlichen papillomavirus Expired - Lifetime EP0357611B1 (de)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
AU559/87 1987-02-26
AUPI055987 1987-02-26
PCT/AU1988/000047 WO1988006634A1 (en) 1987-02-26 1988-02-24 A method of detection of carcinogenic human papillomavirus

Publications (3)

Publication Number Publication Date
EP0357611A1 true EP0357611A1 (de) 1990-03-14
EP0357611A4 EP0357611A4 (en) 1991-10-02
EP0357611B1 EP0357611B1 (de) 1995-04-26

Family

ID=3772035

Family Applications (1)

Application Number Title Priority Date Filing Date
EP88902077A Expired - Lifetime EP0357611B1 (de) 1987-02-26 1988-02-24 Verfahren zum nachweis des karzinogenen menschlichen papillomavirus

Country Status (6)

Country Link
US (2) US5783412A (de)
EP (1) EP0357611B1 (de)
JP (3) JP3096704B2 (de)
AT (1) ATE121794T1 (de)
DE (1) DE3853678T2 (de)
WO (1) WO1988006634A1 (de)

Families Citing this family (45)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5182377A (en) * 1988-09-09 1993-01-26 Hoffmann-La Roche Inc. Probes for detection of human papillomavirus
US5447839A (en) * 1988-09-09 1995-09-05 Hoffmann-La Roche Inc. Detection of human papillomavirus by the polymerase chain reaction
DE3838269A1 (de) * 1988-11-11 1990-05-17 Behringwerke Ag Nachweis humaner papillomavirus dna und ihrer expression in zervix-abstrichen
GB2225112A (en) * 1988-11-22 1990-05-23 Ici Plc Hybridisation probes
US5043272A (en) * 1989-04-27 1991-08-27 Life Technologies, Incorporated Amplification of nucleic acid sequences using oligonucleotides of random sequence as primers
JP2791685B2 (ja) * 1989-06-08 1998-08-27 寳酒造株式会社 パピローマウイルスの検出方法
US5863717A (en) * 1989-11-03 1999-01-26 Abbott Laboratories Use of conserved oligonucleotide primers to amplify human papillomavirus DNA sequences
US5580970A (en) * 1989-12-01 1996-12-03 Amoco Corporation Detection of HPV transcripts
DE69033850T2 (de) * 1989-12-01 2002-06-06 Vysis, Inc. Nukleinsäuresonden zum Nachweis von HPV-Transkripts
JPH05501058A (ja) * 1989-12-04 1993-03-04 アイシス・ファーマシューティカルス・インコーポレーテッド 乳頭腫ウイルス属のアンチセンスオリゴヌクレオチド阻害
NL9000134A (nl) * 1990-01-19 1991-08-16 Stichting Res Fonds Pathologie Primers en werkwijze voor het detecteren van humaan papilloma virus genotypen m.b.v. pcr.
DK0515506T3 (da) * 1990-02-16 2000-05-08 Hoffmann La Roche Fremgangsmåde til påvisning af carcinogene humane papillomavirus
GB9015845D0 (en) * 1990-07-19 1990-09-05 Emery Vincent C Diagnostic method
IL97226A0 (en) * 1991-02-13 1992-05-25 Orgenics Ltd Detection of high and low risk human papillomavirus by enzymatic amplification of dna
US5346811A (en) * 1991-07-22 1994-09-13 Cerveceria Polar Method and products for human papillomavirus detection
US5565339A (en) * 1992-10-08 1996-10-15 Hoffmann-La Roche Inc. Compositions and methods for inhibiting dimerization of primers during storage of polymerase chain reaction reagents
DE4409436A1 (de) * 1994-03-19 1995-09-21 Boehringer Mannheim Gmbh Verfahren zur Bearbeitung von Nukleinsäuren
DE19506561C1 (de) * 1995-02-24 1996-10-10 Deutsches Krebsforsch Verfahren zur Früherkennung von HPV-assoziierten Karzinomen bzw. von hochgradigen, durch HPV-verursachten Dysplasien
DE19526717A1 (de) * 1995-07-21 1997-01-23 Florian Dr Med Heirler Verfahren zur Diagnose des Zervixkarzinoms
AU726047B2 (en) * 1995-11-15 2000-10-26 Gen-Probe Incorporated Nucleic acid probes complementary to human papillomavirus nucleic acid and related methods and kits
US7569344B2 (en) * 1998-10-26 2009-08-04 Ventana Medical Systems, Inc. Detection of human papilloma virus in papanicolaou (Pap) smears
AU1324000A (en) * 1998-10-26 2000-05-15 Ventana Medical Systems, Inc. Detection of human papilloma virus in papanicolaou (pap) smears
CA2365901A1 (en) 1999-04-14 2000-10-19 Musc Foundation For Research Development Tissue-specific and pathogen-specific toxic agents and ribozymes
US20060275784A1 (en) * 1999-10-26 2006-12-07 Ventana Medical Systems, Inc. Detection of Human Papilloma Virus in Papanicolaou (Pap) Smears
US6936443B2 (en) * 2000-04-03 2005-08-30 Cytyc Corporation Detection and typing of human papillomavirus using PNA probes
DE60141205D1 (de) * 2000-04-03 2010-03-18 Cytyc Corp Nachweis und typisierung des papillomavirus mittels pna-sonden
US6811549B2 (en) 2001-02-16 2004-11-02 William H. Fleming Administration of therapeutic or diagnostic agents using interlabial pad
EP2267155B2 (de) * 2002-01-07 2016-09-07 Norchip A/S Detektionsverfahren für Papillomavirus mRNA
US20050100554A1 (en) * 2002-02-14 2005-05-12 Amanda Jackson Complexes and methods of using same
WO2004003199A1 (ja) * 2002-07-01 2004-01-08 Kankyo Engineering Co., Ltd. 核酸若しくは遺伝子の新規取得方法
US20040137551A1 (en) * 2003-01-13 2004-07-15 Markovic Nenad S. Cervical acid phosphatase - papanicolaou (CAP-PAP) test kit, method and accesories, processes for producing and using the same
EP1664348B8 (de) 2003-09-25 2019-06-12 Third Wave Technologies, Inc. Hpv-nachweis
WO2005033333A2 (en) 2003-10-07 2005-04-14 Dako Denmark A/S Methods and compositions for the diagnosis of cancer
EP1819835B1 (de) 2004-12-08 2010-08-04 Gen-Probe Incorporated Nukleinsäuredetektion aus verschiedenen typen des humanen papillomavirus
GB0500999D0 (en) * 2005-01-18 2005-02-23 Glaxosmithkline Biolog Sa Detection method and materials therefor
US20070031826A1 (en) * 2005-08-05 2007-02-08 My Gene Diagnostic kit for determining the genotype of a human papilloma virus and method of using thereof
JPWO2007148762A1 (ja) * 2006-06-22 2009-11-19 雅彦 黒田 ヒトパピローマウイルス（ｈｐｖ）の検出方法、それに用いるプライマーセットおよび検出キット
US8080643B2 (en) * 2006-09-05 2011-12-20 Third Wave Technologies, Inc. HPV primers
US8398606B2 (en) * 2007-02-28 2013-03-19 QuantrRx Biomedical Corporation Folded perineal pad
KR100914911B1 (ko) * 2007-07-10 2009-08-31 의료법인제일의료재단 실시간 중합효소 연쇄반응과 ｈｐｖ ｄｎａ 칩을 이용한정량 및 정성적 인유두종바이러스 검사 방법 및 이를 위한검사키트
KR100906124B1 (ko) 2007-07-19 2009-07-07 캐치바이진 주식회사 고 위험군 인유두종바이러스 ｄｎａ 의 정성 검사 방법 및이의 정성 검사용 키트
DE102008035903A1 (de) * 2008-07-31 2010-02-18 Epcos Ag Elektrisches Bauelement und Verfahren zur Herstellung
NO330943B1 (no) * 2009-04-30 2011-08-22 Unilabs Telelabs As En metode for detektering og/eller typebestemmelse og/eller kvantifisering av human papillomavirus (HPV) type, primere og prober derav samt diagnostisk sett og anvendelse derav.
JP6153866B2 (ja) 2010-05-25 2017-06-28 キアゲン ガイサーズバーグ アイエヌシー． 迅速なハイブリッド捕捉アッセイ、及び関連する戦略的に切断されたプローブ
KR101761701B1 (ko) 2014-07-29 2017-07-26 의료법인 제일의료재단 Hpv 특이적 프로브 및 이를 포함하는 hpv 유전자형 검사용 dna 칩

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES8706823A1 (es) * 1985-03-28 1987-06-16 Cetus Corp Un procedimiento para detectar la presencia o ausencia de al menos una secuencia especifica de acidos nucleicos en una muestra
US5176995A (en) * 1985-03-28 1993-01-05 Hoffmann-La Roche Inc. Detection of viruses by amplification and hybridization
US4683195A (en) * 1986-01-30 1987-07-28 Cetus Corporation Process for amplifying, detecting, and/or-cloning nucleic acid sequences
US4683202A (en) * 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
NL9000134A (nl) * 1990-01-19 1991-08-16 Stichting Res Fonds Pathologie Primers en werkwijze voor het detecteren van humaan papilloma virus genotypen m.b.v. pcr.

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO8806634A1 *

Also Published As

Publication number Publication date
JP2000189200A (ja) 2000-07-11
ATE121794T1 (de) 1995-05-15
DE3853678D1 (de) 1995-06-01
EP0357611A4 (en) 1991-10-02
AU1393888A (en) 1988-09-26
AU611135B2 (en) 1991-06-06
WO1988006634A1 (en) 1988-09-07
JPH02502334A (ja) 1990-08-02
US6218104B1 (en) 2001-04-17
JP2001197894A (ja) 2001-07-24
JP3096704B2 (ja) 2000-10-10
US5783412A (en) 1998-07-21
DE3853678T2 (de) 1995-08-31
EP0357611B1 (de) 1995-04-26

Similar Documents

Publication Publication Date Title
EP0357611B1 (de) Verfahren zum nachweis des karzinogenen menschlichen papillomavirus
Gregoire et al. Amplification of human papillomavirus DNA sequences by using conserved primers
Lungu et al. Typing of human papillomaviruses by polymerase chain reaction amplification with L1 consensus primers and RFLP analysis
Bouda et al. “High risk” HPV types are frequently detected in potentially malignant and malignant oral lesions, but not in normal oral mucosa
Pao et al. Identification of human papillomavirus DNA sequences in peripheral blood mononuclear cells
Soler et al. Detection of human papillomavirus types 6, 11, 16 and 18 in mucosal and cutaneous lesions by the multiplex polymerase chain reaction
Tham et al. Diagnostic Sensitivity of Polymerase Chain Reaction and Southern Blot Hybridization for the Detection df Human Papillomavirus DNA in Biopsy Specimens from Cervical Lesions
Yoshida et al. Quantitative real-time polymerase chain reaction analysis of the type distribution, viral load, and physical status of human papillomavirus in liquid-based cytology samples from cervical lesions
EP3887552A1 (de) Assoziation zwischen der integration von hpv- oder hiv-virusgenomen und der schwere und/oder dem klinischen ergebnis von erkrankungen wie hpv-assoziierten zervikalen läsionen oder aids-pathologie
EP2373819B1 (de) Testverfahren mit mehreren grössen
Miller et al. Human papillomavirus type 16 DNA in esophageal carcinomas from Alaska natives
Pfister Detection of human papillomavirus infection and prospects for diagnostics
JP2007524421A (ja) ヒトパピローマウイルスの検出
Konno et al. Progression of squamous cell carcinoma of the uterine cervix from cervical intraepithelial neoplasia infected with human papillomavirus: a retrospective follow-up study by in situ hybridization and polymerase chain reaction
Park et al. Detection of human papillomavirus genotypes in cervical neoplasia from Korean women using polymerase chain reaction
Herckenrode et al. Prevalence of human papillomavirus (HPV) infection in basque country women using slot‐blot hybridization: A survey of women at low risk of developing cervical cancer
AU611135C (en) A method of detection of carcinogenic human papillomavirus
Bavin et al. Comparison of cervical cytology and the polymerase chain reaction for HPV 16 to identify women with cervical disease in a general practice population
Salam et al. General primer‐mediated polymerase chain reaction for simultaneous detection and typing of human papillomavirus DNA in laryngeal squamous cell carcinomas
Sigurdsson et al. Human papillomavirus (HPV) in an Icelandic population: the role of HPV DNA testing based on hybrid capture and PCR assays among women with screen‐detected abnormal Pap smears
Soler et al. Detection of multiple types of human papillomavirus in a giant condyloma from a grafted patient. Analysis by immunohistochemistry, in situ hybridisation, Southern blot and polymerase chain reaction
Anceschi et al. Multiple primer pairs polymerase chain reaction for the detection of human papillomavirus types
Nakazawa et al. Detection of human papillomavirus types 16 and 18 in the exfoliated cervical cells using the polymerase chain reaction
Park et al. Comparison of L1 consensus primers with E6 type specific primers for detection of human papillomaviruses in paraffin sections of cervical neoplasia
Masafumi et al. Non-isotopic detection of HPV DNA in cervical smears using dot-blot hybridization

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 19890824

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH DE FR GB IT LI LU NL SE

A4 Supplementary search report drawn up and despatched

Effective date: 19910813

AK Designated contracting states

Kind code of ref document: A4

Designated state(s): AT BE CH DE FR GB IT LI LU NL SE

17Q First examination report despatched

Effective date: 19921216

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): AT BE CH DE FR GB IT LI LU NL SE

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: IT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRE;WARNING: LAPSES OF ITALIAN PATENTS WITH EFFECTIVE DATE BEFORE 2007 MAY HAVE OCCURRED AT ANY TIME BEFORE 2007. THE CORRECT EFFECTIVE DATE MAY BE DIFFERENT FROM THE ONE RECORDED.SCRIBED TIME-LIMIT

Effective date: 19950426

Ref country code: FR

Effective date: 19950426

Ref country code: BE

Effective date: 19950426

Ref country code: NL

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 19950426

Ref country code: AT

Effective date: 19950426

REF Corresponds to:

Ref document number: 121794

Country of ref document: AT

Date of ref document: 19950515

Kind code of ref document: T

REF Corresponds to:

Ref document number: 3853678

Country of ref document: DE

Date of ref document: 19950601

EN Fr: translation not filed
NLV1 Nl: lapsed or annulled due to failure to fulfill the requirements of art. 29p and 29m of the patents act
PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LU

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 19960229

PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

26N No opposition filed
REG Reference to a national code

Ref country code: CH

Ref legal event code: PUE

Owner name: THE UNIVERSITY OF SYDNEY TRANSFER- BIOSEARCH INTER

REG Reference to a national code

Ref country code: GB

Ref legal event code: 732E

REG Reference to a national code

Ref country code: CH

Ref legal event code: PFA

Free format text: BIOSEARCH INTERNATIONAL PTY LTD.,LEVEL 10 60 PARK STREET,SYDNEY/NSW 2000 (AU) TRANSFER- BIOSEARCH INTERNATIONAL PTY LTD.,5TH FLOOR 99 ELIZABETH STREET,SYDNEY/NSW 2000 (AU)

REG Reference to a national code

Ref country code: GB

Ref legal event code: IF02

REG Reference to a national code

Ref country code: CH

Ref legal event code: NV

Representative=s name: BUGNION S.A.

Ref country code: CH

Ref legal event code: PUE

Owner name: POLARTECHNICS LTD

Free format text: BIOSEARCH INTERNATIONAL PTY LTD.#5TH FLOOR 99 ELIZABETH STREET#SYDNEY/NSW 2000 (AU) -TRANSFER TO- POLARTECHNICS LTD#LEVEL 1, 140 WILLIAM STREET SYDNEY,#NEW SOUTH WALES 2011 (AU)

REG Reference to a national code

Ref country code: GB

Ref legal event code: 732E

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: CH

Payment date: 20070516

Year of fee payment: 20

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: DE

Payment date: 20070517

Year of fee payment: 20

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: SE

Payment date: 20070529

Year of fee payment: 20

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: GB

Payment date: 20070516

Year of fee payment: 20

REG Reference to a national code

Ref country code: GB

Ref legal event code: PE20

REG Reference to a national code

Ref country code: CH

Ref legal event code: PL

EUG Se: european patent has lapsed
PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: GB

Free format text: LAPSE BECAUSE OF EXPIRATION OF PROTECTION

Effective date: 20080223